The results of early clinical trials using stem cells clearly offer the promise of a new path to addressing a variety of medical problems that range from creating engineered tissues for autologous grafts to repairing a variety of organs. Mesenchymal stem cells (MSCs) are especially attractive candidates for stem cell therapy given their documented characteristic of homing to the target tissue, ability to differentiate into a host of different cell types, and role as a paracrine source thus facilitating the repair and growth of neighboring cells. A notable problem with MSC therapy, however, is the well documented cell death (>50%) that occurs upon isolation, processing and transfusion. This issue has led many groups to "prime" MSC prior to transfusion by transient exposure to stress regimes such as hypoxia. CPSI-Biotech is currently developing a series of culture reagents, CellGuard, that are designed to prevent the loss of cell viability and function of human cells during bioprocessing. This Phase 1 project is designed to develop CellGuard-MSC - a media supplement designed to protect MSC during isolation, processing and transfusion at normothermic temperatures so that both the cell number and differentiation potential are maintained in a more native state. CellGuard is cell and process specific and designed based on the identification and modulation of the key cell stress pathways that are activated in human cells during bioprocessing. Research outlined in this proposal will (1) determine which stress pathways are activated during MSC processing, (2) identify modulators of these pathways and (3) test a prototype CellGuard-MSC for its ability to maintain MSCs under conditions representative of bioprocessing and transfusion. Phase 2 will be focused on additional stem cell types used in cell therapy as well as further development and characterization of CellGuard-MSC. Source-specific MSCs such as adipose-derived, corneal-derived and bone marrow-derived MSCs will be included in the Phase 2 program. The successful completion of this project will result in an enhancement of MSC cell therapy through the development of a molecular "coat of armor" that will protect the integrity and differentiation potential of MSCs during cell isolation and processing. PUBLIC HEALTH RELEVANCE: This project is designed to conduct research that will lead to the development of a series of products, termed CellGuard, which will protect human cells during manipulation. A variety of human cell types are now being used to treat cancer, diabetes, and also serve as the building blocks of engineered tissues used in the biomedical field. Manipulating human cells such that they can be used for these purposes often compromises both cell function and viability. The CellGuard portfolio of products is designed to protect these cells when they are subjected to these harsh conditions on the path for clinical application. This particular project is designed to develop a set of solutions that will protect human mesenchymal stem cells that are currently being used in a variety of stem cell applications ranging from cell therapy for the heart to treating diabetes.